Journal
NATURE STRUCTURAL & MOLECULAR BIOLOGY
Volume 18, Issue 1, Pages 100-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nsmb.1945
Keywords
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Funding
- Howard Hughes Medical Institute
- US National Institute of General Medical Sciences Medical Scientist National Research Service [5 T32 GM07281]
- US National Institutes of Health [R01-GM72688, U54-GM74946, R01-GM61835]
- US Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-06CH11357]
- Michigan Economic Development Corporation
- Michigan Technology Tri-Corridor [085P1000817]
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R37GM061835, R01GM072688, T32GM007281, U54GM074946, R01GM061835, U01GM094588] Funding Source: NIH RePORTER
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RNA crystallization and phasing represent major bottlenecks in RNA structure determination. Seeking to exploit antibody fragments as RNA crystallization chaperones, we have used an arginine-enriched synthetic Fab library displayed on phage to obtain Fabs against the class I ligase ribozyme. We solved the structure of a Fab-ligase complex at 3.1-angstrom resolution using molecular replacement with Fab coordinates, confirming the ribozyme architecture and revealing the chaperone's role in RNA recognition and crystal contacts. The epitope resides in the GAAACAC sequence that caps the P5 helix, and this sequence retains high-affinity Fab binding within the context of other structured RNAs. This portable epitope provides a new RNA crystallization chaperone system that easily can be screened in parallel to the U1A RNA-binding protein, with the advantages of a smaller loop and Fabs' high molecular weight, large surface area and phasing power.
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